Underwater source detection using a spatial stationarity test.
The problem of detecting a source in shallow water is addressed. The complexity of such a propagation channel makes precise modeling practically impossible. This lack of accuracy causes a deterioration in the performance of the optimal detector and motivates the search for suboptimal detectors which are insensitive to uncertainties in the propagation model. A novel, robust detector which measures the degree of spatial stationarity of a received field is presented. It exploits the fact that a signal propagating in a bounded channel induces spatial nonstationarity to a higher degree than mere background noise. The performance of the proposed detector is evaluated using both simulated data and experimental data collected in the Mediterranean Sea. This performance is compared to those of three other detectors, employing different extents of prior information. It is shown that when the propagation channel is not completely known, as is the case of the experimental data, the novel detector outperforms the others in terms of threshold signal-to-noise ratio (SNR). In the presence of environmental mismatch, the threshold SNR of the novel detector for the experimental data appears 2-5 dB lower than the other detectors. That is, this detector couples good performance with robustness to propagation uncertainties.


Radiation-induced neuropathies in head and neck cancer: prevention and treatment modalities.
Head and neck cancer (HNC) is the sixth most common human malignancy with a global incidence of 650,000 cases per year. Radiotherapy (RT) is commonly used as an effective therapy to treat tumours as a definitive or adjuvant treatment. Despite the substantial advances in RT contouring and dosage delivery, patients suffer from various radiation-induced complications, among which are toxicities to the nervous tissues in the head and neck area. Radiation-mediated neuropathies manifest as a result of increased oxidative stress-mediated apoptosis, neuroinflammation and altered cellular function in the nervous tissues. Eventually, molecular damage results in the formation of fibrotic tissues leading to susceptible loss of function of numerous neuronal substructures. Neuropathic sequelae following irradiation in the head and neck area include sensorineural hearing loss, alterations in taste and smell functions along with brachial plexopathy, and cranial nerves palsies. Numerous management options are available to relieve radiation-associated neurotoxicities notwithstanding treatment alternatives that remain restricted with limited benefits. In the scope of this review, we discuss the use of variable management and therapeutic modalities to palliate common radiation-induced neuropathies in head and neck cancers.


Genetic association analyses of PDYN polymorphisms with heroin and cocaine addiction.
Genetic factors are believed to account for 30-50% of the risk for cocaine and heroin addiction. Dynorphin peptides, derived from the prodynorphin (PDYN) precursor, bind to opioid receptors, preferentially the kappa-opioid receptor, and may mediate the aversive effects of drugs of abuse. Dynorphin peptides produce place aversion in animals and produce dysphoria in humans. Cocaine and heroin have both been shown to increase expression of PDYN in brain regions relevant for drug reward and use. Polymorphisms in PDYN are therefore hypothesized to increase risk for addiction to drugs of abuse. In this study, 3 polymorphisms in PDYN (rs1022563, rs910080 and rs1997794) were genotyped in opioid-addicted [248 African Americans (AAs) and 1040 European Americans (EAs)], cocaine-addicted (1248 AAs and 336 EAs) and control individuals (674 AAs and 656 EAs). Sex-specific analyses were also performed as a previous study identified PDYN polymorphisms to be more significantly associated with female opioid addicts. We found rs1022563 to be significantly associated with opioid addiction in EAs [P = 0.03, odds ratio (OR) = 1.31; false discovery rate (FDR) corrected q-value]; however, when we performed female-specific association analyses, the OR increased from 1.31 to 1.51. Increased ORs were observed for rs910080 and rs199774 in female opioid addicts also in EAs. No statistically significant associations were observed with cocaine or opioid addiction in AAs. These data show that polymorphisms in PDYN are associated with opioid addiction in EAs and provide further evidence that these risk variants may be more relevant in females.


Jaw injuries of independence victims from the 1991 War in Croatia.
In the aetiology of maxillofacial injuries, car accidents, violence, sports injuries and lately war injuries are frequently mentioned. The purpose of this study was to exhibit and analyse types of jaw injuries on bodies exhumed from massive and individual graves located in regions temporarily occupied during the War in Croatia that lasted from 1991 to 1995. The sample was the post-mortal documentation of the orofacial region (set of teeth, photographs, radiographic images) of 1068 victims exhumed from massive grave sites in Croatia. The jaw traumatism was analysed on the whole sample as well as on individual graves, whilst the analysis of trauma frequency was performed separately. Descriptive statistics were computed and the value of P < .05 was accepted as statistically significant. Results of this study showed that out of 1068 examined corpses, 332 had midface and lower face fractures, which was 31.1% of the total number. Lower face fractures were more frequent with 28.1%. Age related frequency analysis showed a significant dependency. There were 34.6% of fractures in the under 30 age group, 34.2% in those aged 30-60, while 21.3% of fractures were noted in the over 60 age group. Female bodies had the lowest number of jaw fractures regardless of the osteoporotic changes. The results of this study suggest that younger and middle aged persons were molested more. Jaw fractures suggest ante-mortal molestation. In females, the more likely fracture causes were the falls of the bodies into the graves or body to body hits.


ANALYSIS OF THE EXPERIMENTAL LESION OF CONNECTIVE TISSUE PRODUCED BY A COMPLEX OF C POLYSACCHARIDE FROM GROUP A STREPTOCOCCI. II. INFLUENCE OF AGE AND HYPERSENSITIVITY.
Rabbits presumably sensitized to components of the streptococcal cell by natural contact with cross-reactive microbial antigens in their environment, display distinctive phases of evolving allergic response to an extract of streptococcal cells. Animals under 4 weeks of age show no gross allergic skin reaction; this is followed successively by age periods in which allergic reactions become most prominent at 4 hours, 23 hours, or 2 to 4 days after injection. A relapsing nodular lesion, produced by cell wall components in the cell extract, occurs in neonatal rabbits which display no hypersensitivity, reaches a peak response in young animals during the period of most active growth, and decreases in severity in older rabbits while the degree of hypersensitivity is increasing. Relapses occur with greater frequency in neonatal than in older rabbits. Passive transfer of spleen cells or serum from sensitized adult rabbits has no influence on the nodular reaction in neonatal animals.


The human mitochondrial import receptor, hTom20p, prevents a cryptic matrix targeting sequence from gaining access to the protein translocation machinery.
Yeast Mas70p and NADH cytochrome b5 reductase are bitopic integral proteins of the mitochondrial outer membrane and are inserted into the lipid-bilayer in an Nin-Ccyto orientation via an NH2-terminal signal-anchor sequence. The signal anchor of both proteins is comprised of a short, positively charged domain followed by the predicted transmembrane segment. The positively charged domain is capable of functioning independently as a matrix-targeting signal in yeast mitochondria in vitro but does not support import into mammalian mitochondria (rat or human). Rather, this domain represents a cryptic signal that can direct import into mammalian mitochondria only if proximal components of the outer membrane import machinery are removed. This can be accomplished either by treating the surface of the intact mitochondria with trypsin or by generating mitoplasts. The import receptor Tom20p (Mas20p/MOM19) is responsible for excluding the cryptic matrix-targeting signal from mammalian mitochondria since replacement of yeast Tom20p with the human receptor confers this property to the yeast organelle while at the same time maintaining import of other proteins. In addition to contributing to positive recognition of precursor proteins, therefore, the results suggest that hTom20p may also have the ability to screen potential matrix-targeting sequences and exclude certain proteins that would otherwise be recognized and imported by distal components of the outer and inner membrane protein-translocation machinery. These findings also indicate, however, that cryptic signals, if they exist within otherwise native precursor proteins, may remain topogenically silent until the precursor successfully clears hTom20p, at which time the activity of the cryptic signal is manifested and can contribute to subsequent translocation and sorting of the polypeptide.


Targeting Leukemia-Initiating Cells in Acute Lymphoblastic Leukemia.
The concept that different leukemias are developmentally distinct and, like in normal hematopoiesis, generated by restricted populations of cells named leukemia-initiating cells (LIC), is becoming more established. These cancer stem-like cells have been assumed to have unique properties, including the capability of self-renewing and giving rise to "differentiated" or non-LICs that make up the whole tumor. Cell populations enriched with LIC activity have been characterized in different hematopoietic malignancies, including human acute lymphoblastic leukemia (ALL). Related studies have also demonstrated that LICs are functionally distinct from bulk cells and modulated by distinct molecular signaling pathways and epigenetic mechanisms. Here we review several biological and clinical aspects related to LICs in ALL, including (i) immunophenotypic characterization of LIC-enriched subsets in human and mouse models of ALL, (ii) emerging therapeutics against regulatory signaling pathways involved in LIC progression and maintenance in T- and B-cell leukemias, (iii) novel epigenetic and age-related mechanisms of LIC propagation, and (iv) ongoing efforts in immunotherapy to eradicate LIC-enriched cell subsets in relapsed and refractory ALL cases. Current conventional treatments do not efficiently eliminate LICs. Therefore, innovative therapeutics that exclusively target LICs hold great promise for developing an effective cure for ALL.


Elevated serum digoxin level associated with coadministered fluoxetine.
After her daughter died, depression developed in a 93-year-old woman with congestive heart failure receiving digoxin therapy. After treatment with fluoxetine hydrochloride she became anorexic and her serum digoxin level increased. A possible interaction, which has not been reported in the literature, may have occurred between fluoxetine and digoxin.


Managing public services in the 1990s. President's lecture. Keele University, MBA (Health Executive) Alumni Association.
The management of public services in the 1990s raises a number of central issues which require to be resolved. On the one hand, efficiency and effectiveness require attention, as in the 1980s or indeed anytime. On the other hand, the distinctive nature of public (and governmental) services needs to be recognized or reasserted. This Keynote Address to the Keele University, Centre for Health Planning and Management MBA (Health Executive) Programme Alumni Association by Howard Davies, Director General of the Confederation of British Industry, sets out some key principles based on the 'British Debate' which are nevertheless generalizable in a number of cases to a wholly international audience.


Low-dose dopamine agonist administration blocks vascular endothelial growth factor (VEGF)-mediated vascular hyperpermeability without altering VEGF receptor 2-dependent luteal angiogenesis in a rat ovarian hyperstimulation model.
No specific treatment is available for ovarian hyperstimulation syndrome (OHSS), the most important complication in infertile women treated with gonadotropins. OHSS is caused by increased vascular permeability (VP) through ovarian hypersecretion of vascular endothelial growth factor (VEGF)-activating VEGF receptor 2 (VEGFR-2). We previously demonstrated in an OHSS rodent model that increased VP was prevented by inactivating VEGFR-2 with a receptor antagonist (SU5416). However, due to its toxicity (thromboembolism) and disruption of VEGFR-2-dependent angiogenic processes critical for pregnancy, this kind of compound cannot be used clinically to prevent OHSS. Dopamine receptor 2 (Dp-r2) agonists, used in the treatment of human hyperprolactinemia including pregnancy, inhibit VEGFR-2-dependent VP and angiogenesis when administered at high doses in animal cancer models. To test whether VEGFR-2-dependent VP and angiogenesis could be segregated in a dose-dependent fashion with the Dp-r2 agonist cabergoline, a well-established OHSS rat model supplemented with prolactin was used. A 100 microg/kg low-dose Dp-r2 agonist cabergoline reversed VEGFR-2-dependent VP without affecting luteal angiogenesis through partial inhibition of ovarian VEGFR-2 phosphorylation levels. No luteolytic effects (serum progesterone levels and luteal apoptosis unaffected) were observed. Cabergoline administration also did not affect VEGF/VEGFR-2 ovarian mRNA levels. Results in the animal model and the safe clinical profile of Dp-r2 agonists encouraged us to administer cabergoline to oocyte donors at high risk for developing the syndrome. Prophylactic administration of cabergoline (5-10 microg/kg x d) decreased the occurrence of OHSS from 65% (controls) to 25% (treatment). Therefore, a specific, safe treatment for OHSS is now available.


Current Understanding of Inflammatory Responses in Acute Kidney Injury.
Acute kidney injury has been a tough complex with increased mortality and morbidity. Inflammatory responses, including innate and adaptive immune responses, involve in the initiation and development of acute kidney injury, especially under the ischemic circumstances. Tubular cells and distinct immune cell subgroups play a critical role in the pathogenesis of inflammation. Current gene therapies show their benefits in renal repair. Here, we reviewed the renal inflammatory infiltration, inflammatory mediators, oxidative stress and potential signaling pathways, which give rise to the kidney diseases, in the mechanism of acute kidney injury. Recent studies showed diverse insights in understanding the pathophysiological process of inflammation related renal injury and provided novel clinical targets for ameliorating acute kidney injury by balancing the facilitation of repairing and prevention of impairing. Interestingly, antisense oligonucleotides of target sequence, local electransfering on solid organ and adenovirus-mediated certain gene overexpression have been promising strategies in alleviating acute kidney injury.


[Effect of unilateral right hemisphere lesions upon recognition of faces tachistoscopically presented to the left hemisphere].
Normal subjects recognize a tachistoscopically presented face faster when it is flashed to the left visual field (right hemisphere) than when it is presented to the right visual field (left hemisphere). In order to assess the capacity of the left hemisphere in processing physiognomical material, a comparison was done between the choice-reaction times of a group of normals and of a group of right brain-damaged patients, to face photographs tachistoscopically projected to the right visual filed (left hemisphere). All subjects were right-handed adult males. In the brain damaged group it took almost as twice as the time taken in the controls for the left hemisphere to perform the task, even when unspecific factors, lengthening the reaction times, were excluded. These results are interpreted as favouring the hypothesis of an "absolute" right hemisphere dominance for recognizing faces, suggesting that in normal conditions a face projected to the left hemisphere is analyzed either exclusively by the right hemisphere or with a preponderant cooperation of it after the transfer of information across commissural pathways.


Variation in the heat shock response and its implication for predicting the effect of global climate change on species' biogeographical distribution ranges and metabolic costs.
The preferential synthesis of heat shock proteins (Hsps) in response to thermal stress [the heat shock response (HSR)] has been shown to vary in species that occupy different thermal environments. A survey of case studies of aquatic (mostly marine) organisms occupying stable thermal environments at all latitudes, from polar to tropical, shows that they do not in general respond to heat stress with an inducible HSR. Organisms that occupy highly variable thermal environments (variations up to >20 degrees C), like the intertidal zone, induce the HSR frequently and within the range of body temperatures they normally experience, suggesting that the response is part of their biochemical strategy to occupy this thermal niche. The highest temperatures at which these organisms can synthesize Hsps are only a few degrees Celsius higher than the highest body temperatures they experience. Thus, they live close to their thermal limits and any further increase in temperature is probably going to push them beyond those limits. In comparison, organisms occupying moderately variable thermal environments (<10 degrees C), like the subtidal zone, activate the HSR at temperatures above those they normally experience in their habitats. They have a wider temperature range above their body temperature range over which they can synthesize Hsps. Contrary to our expectations, species from highly (in comparison with moderately) variable thermal environments have a limited acclimatory plasticity. Due to this variation in the HSR, species from stable and highly variable environments are likely to be more affected by climate change than species from moderately variable environments.


[Differential diagnosis of the hypodynamic syndrome in myocardial infarct and congestive heart failure].
The clinical course of myocardial infarction (MI), central hemodynamics, cardiac contractility, gas exchange and blood lactate and pyruvate levels were examined in 169 MI patients with hypodynamic circulation. Group 1 was made up of 23 patients whose low cardiac output and heart performance were not accompanied with tachycardia, increased double product and clinical signs of congestive heart failure (CHF), i.e. patients with the hypodynamic syndrome. Group 2 was composed of 146 patients with signs of second- or third-degree CHF (by M. J. Walk's classification). Unlike the patients with the hypodynamic syndrome those with CHF showed reduced cardiac output and heart performance that did not correspond to metabolic requirements of the body as evidenced by increased blood lactate levels and markedly disturbed gas exchange. The patients with the hypodynamic syndrome demonstrated a favorable course of MI and needed no active hemodynamic correction.


Telemedicine and the law.
The delivery of medical services through the use of modern technology is proving of significant benefit, particularly in remote communities where transportation of patients and medical practitioners is problematic. Technical systems supporting telemedicine-based consultations have been available for some years and more recently diagnostic services such as pathology and radiology have embraced this new technology. While teleradiology and telepathology allow medical services to be delivered across vast distances from highly populated areas to regions of low population density, the same technology can also deliver services across national and international boundaries. Where the patient and medical practitioner are located in different jurisdictions, issues arise regarding the regulation of medical service provision and this has an impact on the registration of medical practitioners, mutual recognition of training and specialisation and the administration of medical service provision. The increasing specialisation of medical practice and manpower shortages in key practice areas would suggest that there will be ongoing expansion of telemedicine services in the years to come. This will require greater standardisation of medical education, together with an improvement in arrangements for mutual recognition of medical practitioners across national boundaries.


Clinical review: scoring systems in the critically ill.
General illness severity scores are widely used in the ICU to predict outcome, characterize disease severity and degree of organ dysfunction, and assess resource use. In this article we review the most commonly used scoring systems in each of these three groups. We examine the history of the development of the initial major systems in each group, discuss the construction of subsequent versions, and, when available, provide recent comparative data regarding their performance. Importantly, the different types of scores should be seen as complementary, rather than competitive and mutually exclusive. It is possible that their combined use could provide a more accurate indication of disease severity and prognosis. All these scoring systems will need to be updated with time as ICU populations change and new diagnostic, therapeutic and prognostic techniques become available.


Measuring the quality of a childhood cancer care delivery system: assessing stakeholder agreement.
We described previously the development of a set of quality indicators (QIs) of a childhood cancer system in Ontario, Canada. The purpose of this study was to determine the acceptability of the proposed set of QIs among stakeholders of the childhood cancer system. A modified Delphi method was used to assess stakeholder agreement on the value of the proposed QIs. A QI evaluation survey was mailed to a stakeholder group of 23 multidisciplinary health care providers, survivors, parents, and policymakers who rated each QI on specific criteria. Prior to an in-person consensus meeting, the distribution of scores was provided to panel members. At the meeting, QIs were reevaluated and discussed in three successive rounds. QIs with 80% or more of panel agreement were considered endorsed. Overall, 20 QIs were endorsed by the panel, measuring all seven quality dimensions of Ontario's Cancer System Quality Index framework. Five QIs were endorsed by 100% of the panel as follows: Five-year event-free survival, chemotherapy admission delay, drug availability, sufficient multidisciplinary staff, and parent satisfaction. Although none of the QIs relating to end-of-life or Satellite care were endorsed, panel members emphasized the need to measure these components of the system. Standardized implementation of the 20 pediatric cancer QIs endorsed by the multidisciplinary stakeholder panel will provide ongoing monitoring of various dimensions of system quality and the development of benchmarks over time, greatly augmenting the ability to identify needed system improvements across populations and jurisdictions.


Membrane interactions and metal ion effects on bilayer permeation of the lipophilic ion modulator DP-109.
DP-109, a lipophilic bivalent metal ion modulator currently under preclinical development for neurodegenerative disorders, was designed to have membrane-associated activity, thereby restricting its action to the vicinity of cell membranes. We describe the application of a colorimetric phospholipid/polydiacetylene (PDA) biomimetic membrane assay in elucidating DP-109 membrane interactions and penetration into lipid bilayers. In this membrane model, visible quantifiable color changes were monitored in studying membrane interactions. The colorimetric data identified a biphasic concentration-dependent interaction, with a break point around the critical micelle concentration (CMC) of DP-109. The kinetics and colorimetric dose-response profile of DP-109 indicate that the compound inserts into the lipid bilayers rather than being localized at the bilayer surface. Analysis of interactions of DP-109 with phospholipid/PDA vesicles in which ionic gradients were imposed indicates that membrane activity of DP-109 is strongly affected by electrochemical gradients imposed by K+ and Zn2+. The ionic gradient effects suggest that the insertion of DP-109 into the membrane may depend on the membrane potential.


A TCTN2 mutation defines a novel Meckel Gruber syndrome locus.
Meckel Gruber syndrome (MKS) is an autosomal recessive multisystem disorder that represents a severe form of ciliopathy in humans and is characterized by significant genetic heterogeneity. In this article, we describe the identification of a novel MKS locus MKS8 that we map to TCTN2, in a multiplex consanguineous family. TCTN2 is a paralog of the recently identified Tectonic 1, which has been shown to modulate sonic hedgehog signaling. Expression analysis at different developmental stages of the murine ortholog revealed a spatial and temporal pattern consistent with the MKS phenotype observed in our patient. The exclusion of this and the other seven MKS genes in our collection of consanguineous Arab MKS families confirms the existence of two additional MKS loci.


Winner of the Brazier Award. The dysmature EEG pattern in infants with bronchopulmonary dysplasia and its prognostic implications.
Bronchopulmonary dysplasia (BPD) is a chronic lung disease which occurs in premature infants who require prolonged mechanical ventilation and supplemental oxygen support. Infants with severe BPD often have neurological sequelae or early mortality. We have observed a pattern of delay in the maturation of cerebral electrical activity of some infants who have more severe forms of BPD. This delayed electroencephalographic (EEG) maturation or 'dysmaturity' is best recognized when the conceptional age (CA) of the premature infant approaches term and is characterized by the presence of EEG activity which normally disappears in premature infants by 36-37 weeks CA. We studied 36 infants with BPD who had serial EEGs and at least 1 EEG between 37 and 42 weeks CA. Dysmature EEGs were found in 23 infants. Unfavorable outcomes were found in 83% (19/23) of these infants, including 22% (5/23) mortality, and 61% (14/23) abnormal or suspect neurological outcome. Thirteen infants had EEGs which lacked dysmature features. Of these infants, 62% (8/13) had normal neurological outcome, and 38% (5/13) had unfavorable outcome (1 death, 2 abnormal, and 2 suspect). In 2 of the 5 infants with unfavorable outcomes without dysmature EEGs, a suppression-burst pattern (known to be associated with neurological sequelae) was found. The presence of dysmature patterns in the near-term or term EEG was associated with significantly less favorable outcome (P = 0.01). Several other clinical factors which were associated with dysmature EEGs are also discussed.


Hypomanic symptoms in female undergraduate students diagnosed with unipolar depression based on scores on the hypomania checklist.
A number of studies have documented high levels of hypomanic symptoms in those diagnosed with depression, suggesting a potential misdiagnosis of bipolar disorder as unipolar depression. Research suggests that undergraduate students have high levels of depression, but whether such misdiagnosis occurs in this population has not been examined. The aim of this study was therefore to examine levels of hypomania in undergraduate students reporting diagnosed depression. An international sample of undergraduate students completed the 32-item Hypomania Checklist (HCL-32). A cohort was analysed for this study, consisting of female undergraduate students reporting a formal diagnosis of depression (n=28). Participants scored high on the HCL-32, with a mean total score of 19.9 (SD=5.4) out of 32. Overall, 85.7% (n=24) scored equal to or above the original cut off point of 14 suggested for bipolar II disorder. Two possible conclusions are suggested by this study. Firstly, there are high levels of hypomanic symptoms in undergraduate students diagnosed with depression, suggesting that a formal diagnosis of bipolar disorder should be pursued in those with high scores. Alternatively, the cut-off points previously suggested for the HCL-32 may not be accurate for use with undergraduate students.


Analysis of a glycinergic inhibitory pathway in the cat retina.
Incubation of cat retinas with 3H-glycine in vitro, followed by horizontal sectioning and autoradiography, showed labeling of 10-12% of bipolar cells and 45% of amacrine cells. To ascertain the effects of glycine-accumulating bipolar and amacrine cells on the response properties of retinal ganglion cells, in vivo iontophoretic studies were performed in the cat eye. Glycine inhibited all ganglion cells, and this action was blocked by strychnine. Aminophosphonobutyric acid (APB) suppressed ON-ganglion cells, but activated OFF-ganglion cells. The influence of APB upon OFF-ganglion cells could be completely blocked by strychnine. In the mudpuppy, APB suppressed ON-bipolar cells without affecting OFF-bipolar cells and without direct effects on other retinal neurons (Slaughter and Miller, 1981, 1985). If APB has the same mode of action in the cat retina, the present results suggest a glycinergic inhibitory input of the ON-channel upon OFF-ganglion cells.


An assessment of the potential miscalibration of cardiovascular disease risk predictions caused by a secular trend in cardiovascular disease in England.
A downwards secular trend in the incidence of cardiovascular disease (CVD) in England was identified through previous work and the literature. Risk prediction models for primary prevention of CVD do not model this secular trend, this could result in over prediction of risk for individuals in the present day. We evaluate the effects of modelling this secular trend, and also assess whether it is driven by an increase in statin use during follow up. We derived a cohort of patients (1998-2015) eligible for cardiovascular risk prediction from the Clinical Practice Research Datalink with linked hospitalisation and mortality records (N = 3,855,660). Patients were split into development and validation cohort based on their cohort entry date (before/after 2010). The calibration of a CVD risk prediction model developed in the development cohort was tested in the validation cohort. The calibration was also assessed after modelling the secular trend. Finally, the presence of the secular trend was evaluated under a marginal structural model framework, where the effect of statin treatment during follow up is adjusted for. Substantial over prediction of risks in the validation cohort was found when not modelling the secular trend. This miscalibration could be minimised if one was to explicitly model the secular trend. The reduction in risk in the validation cohort when introducing the secular trend was 35.68 and 33.24% in the female and male cohorts respectively. Under the marginal structural model framework, the reductions were 33.31 and 32.67% respectively, indicating increasing statin use during follow up is not the only the cause of the secular trend. Inclusion of the secular trend into the model substantially changed the CVD risk predictions. Models that are being used in clinical practice in the UK do not model secular trend and may thus overestimate the risks, possibly leading to patients being treated unnecessarily. Wider discussion around the modelling of secular trends in a risk prediction framework is needed.


Impact of intensive glycemic control on the incidence of atrial fibrillation and associated cardiovascular outcomes in patients with type 2 diabetes mellitus (from the Action to Control Cardiovascular Risk in Diabetes Study).
Atrial fibrillation (AF) is prevalent in patients with type 2 diabetes mellitus (DM) and is associated with markers of poor glycemic control; however, the impact of glycemic control on incident AF and outcomes is unknown. The aims of this study were to prospectively evaluate if intensive glycemic control in patients with DM affects incident AF and to evaluate morbidity and mortality in patients with DM and incident AF. A total of 10,082 patients with DM from the Action to Control Cardiovascular Risk in Diabetes (ACCORD) cohort were studied in a randomized, double-blind fashion. Participants were randomized to an intensive therapeutic strategy targeting a glycated hemoglobin level of <6.0% or a standard strategy targeting a glycated hemoglobin level of 7.0% to 7.9%. Incident AF occurred in 159 patients (1.58%) over the follow-up period, at a rate of 5.9 per 1,000 patient-years in the intensive-therapy group and a rate of 6.37 per 1,000 patient-years in the standard-therapy group (p = 0.52). In a multivariate model, predictors of incident AF were age, weight, diastolic blood pressure, heart rate, and heart failure history. Patients with DM and new-onset AF had a hazard ratio of 2.65 for all-cause mortality (95% confidence interval 1.8 to 3.86, p <0.0001), a hazard ratio of 2.1 for myocardial infarction (95% confidence interval 1.33 to 3.31, p = 0.0015), and a hazard ratio of 3.80 for the development of heart failure (95% confidence interval 2.48 to 5.84, p <0.0001). In conclusion, intensive glycemic control did not affect the rate of new-onset AF. Patients with DM and incident AF had an increased risk for morbidity and mortality compared with those without AF.


Three-dimensional visualization and analysis of flowing droplets in microchannels using real-time quantitative phase microscopy.
Recent years have witnessed the development of droplet-based microfluidics as a useful and effective tool for high-throughput analysis in biological, chemical and environmental sciences. Despite the flourishing development of droplet manipulation techniques, only a few methods allow for label-free and quantitative inspection of flowing droplets in microchannels in real-time and in three dimensions (3-D). In this work, we propose and demonstrate the application of a real-time quantitative phase microscopy (RT-QPM) technique for 3-D visualization of droplets, and also for full-field and label-free measurement of analyte concentration distribution in the droplets. The phase imaging system consists of a linear-CCD-based holographic microscopy configuration and an optofluidic phase-shifting element, which can be used for retrieving quantitative phase maps of flowing objects in the microchannels with a temporal resolution only limited to the frame rate of the CCD camera. To demonstrate the capabilities of the proposed imaging technique, we have experimentally validated the 3-D image reconstruction of the droplets generated in squeezing and dripping regimes and quantitatively investigated the volumetric and morphological variation of droplets as well as droplet parameters related to the depth direction under different flow conditions. We also demonstrated the feasibility of using this technique, as a refractive index sensor, for in-line quantitative measurement of carbamide analyte concentration within the flowing droplets.


Toxic responses to deltamethrin (DM) low doses on gonads, sex hormones and lipoperoxidation in male rats following subcutaneous treatments.
Deltamethrin (DM) is a alpha- cyano pyrethroid insecticide used extensively in pest control. Although initially thought to be least toxic, a number of recent reports showed its toxicity in mammalian and non-mammalian laboratory and wildlife animal species. The present study, carried out in male rats, is a contribution to explore some mechanisms underlying DM toxicity. The aim of the present research was to investigate the effect of different subcutaneous treatments with DM (2 ppm for 30 days, 20 ppm for 45 days and 200 ppm for 60 days) on testes histopathology, sex hormones and oxidative stress from male rats. Our study mentioned an hypospermatogenesis within the testes accompanied by some apoptotic figures in particular cell fragments into the seminiferous tubules (ST)' lumen. The results obtained show that follicule-stimulating hormone (FSH), luteinizing hormone (LH) and testosterone can be differently influenced in rats. In fact, findings of the present investigation mention a significant decrease (p < or = 0.05) of FSH, LH and testosterone at the highest DM dose. Whereas a significant reduction of FSH was noticed after 45 days of treatment. The assessment of oxidative stress by malondialdehyde (MDA) measurements in plasma revealed a significant increase of this parameter after 30 days, 45 days or 60 days. In conclusion the study shows that subcutaneous DM treatment produces an arrest of spermatogenesis, a significant disharmony in sex hormones and MDA levels in rats that is related to dose, length of treatment and to the lipid peroxidation which may be one of the molecular mechanisms involved in DM-induced gonads toxicity.


Fourth report of the cooperative, open-ended study of slowly growing mycobacteria by the International Working Group on Mycobacterial Taxonomy.
The open-ended study of the International Working Group on Mycobacterial Taxonomy is an ongoing project to characterize slowly growing strains of mycobacteria that do not belong to well-established or thoroughly characterized species. In this fourth report we describe two numerical taxonomic clusters that represent subspecies or biovars of Mycobacterium simiae, one cluster that encompasses the erstwhile type strain of the presently invalid species "Mycobacterium paraffinicum," one cluster that is phenotypically very similar to Mycobacterium avium and Mycobacterium intracellulare but may be a separate genospecies, one cluster that appears to be phenotypically distinct from M. avium but reacts with a nucleic acid probe specific for M. avium, and three tentatively defined clusters in proximity to a cluster that encompasses the type strain of Mycobacterium malmoense. Of special practical interest is the fact that one of the latter three clusters is composed of clinically significant scotochromogenic bacteria that can be misidentified as the nonpathogenic organism Mycobacterium gordonae if insufficient biochemical tests are performed.


Supraphysiological testosterone enanthate administration prevents bone loss and augments bone strength in gonadectomized male and female rats.
High-dose testosterone enanthate (TE) may prevent hypogonadism-induced osteopenia. For this study, 3-mo-old male and female Fisher SAS rats underwent sham surgery, gonadectomy (GX), or GX plus 28 days TE administration (7.0 mg/wk). GX reduced serum sex hormones (i.e., testosterone, dihydrotestosterone, and estradiol) (P < 0.05) in both sexes and bone concentrations of testosterone (males only), and estradiol (females only). GX also elevated urine deoxypyridinoline/creatinine in both sexes and serum osteocalcin (females only), findings that are consistent with high-turnover osteopenia. GX reduced cancellous bone volume (CBV) and increased osteoid surfaces in tibia of both sexes. GX males also experienced reduced trabecular number and width and increased trabecular separation, whereas GX females experienced increased osteoblast and osteoid surfaces. Bone biomechanical characteristics remained unaffected by GX, except that femoral stiffness was reduced in females. In contrast, TE administration to GX rats elevated serum and bone androgens to supraphysiological concentrations in both sexes but altered neither serum nor bone estradiol in males. Additionally, TE did not prevent GX-induced reductions in serum or bone estradiol in females. TE also reduced markers of high-turnover osteopenia in both sexes. In males, TE prevented GX-induced changes in trabecular number and separation, CBV, and osteoid surfaces while diminishing osteoblast and osteoclast surfaces; however, these changes were not fully prevented in females. In both sexes, TE increased femoral length and femoral maximal strength to above that of Sham and GX animals while preventing the loss of femoral stiffness in females. In conclusion, TE administration appears protective of cancellous bone in male rats and augments cortical bone strength in both sexes.


Parathyroid hormone [1-34] improves articular cartilage surface architecture and integration and subchondral bone reconstitution in osteochondral defects in vivo.
The 1-34 amino acid segment of the parathyroid hormone (PTH [1-34]) mediates anabolic effects in chondrocytes and osteocytes. The aim of this study was to investigate whether systemic application of PTH [1-34] improves the repair of non-osteoarthritic, focal osteochondral defects in vivo. Standardized cylindrical osteochondral defects were bilaterally created in the femoral trochlea of rabbits (n = 8). Daily subcutaneous injections of 10 μg PTH [1-34]/kg were given to the treatment group (n = 4) for 6 weeks, controls (n = 4) received saline. Articular cartilage repair was evaluated by macroscopic, biochemical, histological and immunohistochemical analyses. Reconstitution of the subchondral bone was assessed by micro-computed tomography. Effects of PTH [1-34] on synovial membrane, apoptosis, and expression of the PTH receptor (PTH1R) were determined. Systemic PTH [1-34] increased PTH1R expression on both, chondrocytes and osteocytes within the repair tissue. PTH [1-34] ameliorated the macro- and microscopic aspect of the cartilaginous repair tissue. It also enhanced the thickness of the subchondral bone plate and the microarchitecture of the subarticular spongiosa within the defects. No significant correlations were established between these coexistent processes. Apoptotic levels, synovial membrane, biochemical composition of the repair tissue, and type-I/II collagen immunoreactivity remained unaffected. PTH [1-34] emerges as a promising agent in the treatment of focal osteochondral defects as its systemic administration simultaneously stimulates articular cartilage and subchondral bone repair. Importantly, both time-dependent mechanisms of repair did not correlate significantly at this early time point and need to be followed over prolonged observation periods.


[Frequency and correlative relation of diagnostic criteria in animal dander allergy].
355 out-patients with signs of bronchial asthma were studied with special reference to animal dander sensitization. Case histories allowed the clinical diagnosis of an allergy to animal dander in 75 patients (21.1%). Skin testing is a useful means, as large reactions appear in most cases of animal dander allergy (60.7% are greater than or equal to + + + -reactions). Skin reactions, however, often are "false positive" or "false negative". Therefore, bronchial provocation tests are necessary if the case history is doubtful and/or the skin reaction is less than a + + + -reaction. There is a positive correlation between skin reaction and bronchial provocation test. No negative inhalation tests were seen when the skin reaction was greater than + + +. However, a positive inhalation test can be correlated with a negative or weakly positive skin test. The probability of common structural properties of different mammalian dander allergens is discussed. Avoidance of exposure to the animal is recommended as the therapy of choice.


Feasibility of standardized methods to specify behavioral pediatric obesity prevention interventions.
Standardized methods are needed to evaluate what occurs within the 'black box' of behavioral interventions to prevent pediatric obesity. The purpose of this research is to evaluate methods to specify the behavior change techniques used and the amount of time spent discussing target weight-related behaviors in an intervention for parents of children at risk for becoming overweight or obese. Independent coders were trained to identify behavior change techniques and time spent discussing weight-related behaviors in audio recordings and transcripts of intervention sessions from 100 randomly selected participants. The behavior change technique taxonomy (BCTTv1) was used to code techniques present in sessions. A newly-developed tool was used to code time spent discussing each target weight-related behavior (e.g., physical activity, screen time). Sessions from a subset of these participants (N = 20) were double coded to evaluate inter-rater reliability. After revisions to coding protocols, coders reliably coded behavior change techniques used and time spent discussing target weight-related behaviors in sessions from the subset of 20 participants. The most commonly discussed target weight-related behavior was physical activity followed by energy intake and fruit and vegetable intake. On average, 13.9 (SD = 2.8) unique behavior change techniques were present across sessions for a given participant. These results offer reliable methods for systematically identifying behavior change techniques used and time spent discussing weight-related behaviors in a pediatric obesity prevention intervention. This work paves the way for future research to identify which specific target behaviors and techniques are most associated with the prevention of unhealthy weight gain in children.


The role of uric acid in mineral bone disorders in chronic kidney disease.
Increasing survival in the chronic kidney disease (CKD) population exposes the bone to the cumulative detrimental sequelae of CKD, now defined physiologically and histopathologically as chronic kidney disease mineral bone disorder (CKD-BMD). This disorder is increasingly recognized as a "nontraditional" driver of morbidity and mortality and presents an opportunity to improve CKD outcomes via research. However, recent advances in the literature on this topic have not yet been collected into a single review. Therefore, this report aims to discuss the disordered renal-bone axis in CKD-BMD, molecular and hormonal drivers, novel treatment strategies, and forthcoming research in a clinician-directed format. A key novel topic will be the unique impact of uric acid on CKD-BMD, which is poised to apply extensive existing research in the uric acid domain to benefit the CKD-BMD population.


Smoking cessation activities by general practitioners and practice nurses.
To assess general practitioners' and practice nurses' self reported behaviour, attitudes, and knowledge in relation to smoking cessation. Two postal surveys of random national samples of 303 GPs (survey 1) and 459 practice nurses (survey 2) covering England and Wales; effective response rates were 75% and 96%, respectively. Survey 1 found that 96% of GPs accepted that intervening against smoking was part of their role and almost all (99%) said that they recorded smoking status when patients registered; 57% reported that they routinely updated their records on smoking status, 50% said they advised smokers to stop during most or all consultations, and 76% said they advised smokers to cut down if they cannot stop. A large majority (83%) said they either recommended or prescribed nicotine replacement therapy (NRT). Although most GPs (86%) thought that NRTs were effective, only a minority thought they were worth the cost (47%) or should be on National Health Service (NHS) prescription (32%). There was little evidence that previous training in smoking cessation was associated with more activity, more positive attitudes, or greater knowledge. Survey 2 found that almost all practice nurses (99%) agreed that intervening against smoking was part of their role and 95% said they advised patients to stop at least occasionally; 71% said they advised smokers to stop at most or all consultations. A majority (74%) said that they recommended NRT to their patients. As with the GPs most practice nurses thought that nicotine replacement was effective (79%), but fewer (42%) thought the cost was justified, and only about half (53%) thought it should be available on NHS prescription. Nurses who said they had been trained in smoking cessation engaged in more activity relating to smoking cessation, had more positive attitudes, and were more knowledgeable. GPs and practice nurses accepted that intervening with smoking was an important part of their role and a large majority reported that they intervened at least with some smokers. This represents a promising baseline from which to proceed in terms of implementation of the new smoking cessation guidelines, but it is hoped that improvements can be made in terms of the frequency of updating records and intervening, and acceptance of the cost-effectiveness of NRT as a life preserving intervention.


Chloroquine prophylaxis, iron/folic-acid supplementation or case management of malaria attacks in primigravidae in western Uganda: effects on congenital malaria and infant haemoglobin concentrations.
A randomized, double-blind, placebo-controlled trial, which compared the effects of three interventions (weekly chloroquine prophylaxis, daily iron and weekly folic-acid supplementation, and case management of malaria) on congenital malaria, maternal haemoglobin (Hb) and foetal outcome, was conducted among primigravidae resident in Hoima district, Uganda. Among 473 babies examined at birth or within 7 days of birth, 198 (42%) were parasitaemic, the level of parasitaemia in an infant being strongly correlated with those of placental (P< 0.01) and maternal, peripheral parasitaemia (P < 0.01). However, 33 (17%) of the parasitaemic babies were born to mothers who had placental but not peripheral parasitaemia, 22 (11%) to mothers who had peripheral but not placental parasitaemia, and 12 (6%) to mothers with neither peripheral nor placental parasitaemia. Overall, 163 babies were each examined for malarial parasites at birth and 1 month later. Of the 76 (47%) found to have parasitaemia at birth, 37 (23%) appeared aparasitaemic at the 1-month follow-up but 28 (17%) were still parasitaemic at that time. Among the babies born to the mothers who only received case management of malaria during pregnancy, parasitaemia at birth was associated with infant anaemia at birth (i.e. < 140 g Hb/litre; P = 0.03). Infants found to be parasitaemic at the 1-month follow-up had lower mean concentrations of Hb at that time than their aparasitaemic counterparts (P= 0.03). Parasitaemia at birth was not significantly associated with low birthweight, in any of three intervention groups. The intervention given to the mother had no significant effect on the parasitaemia of her baby, either at birth or at the age of 1 month. Congenital malaria per se may have little influence on birthweight but may have an impact on infant anaemia. In conclusion, congenital parasitaemia was not associated with birthweight, but was related to anaemia at birth in infants born to women who had only received active case management during their pregnancies.


Orthotopic liver transplantation in U.S. veterans under primary tacrolimus immunosupression.
The evolution and refinement of surgical techniques, per ioperative patient care, and immunosuppression hav~ estab~ished orthoto~ic li~er transplantation (OLTX) as a ~ighly successful therapeutic modality for patients wrth end-stage hver disease. In February 1989,Tacrohmus (Prograf®, formerly FK 506)was first used successfully at the University of Pittsburgh Medical Center to treat patients with rejection refractory to cyclosporine-based immunosuppression." Clinical trials utilizing Tacrolimus in solid organ transplantation followed, and in April of 1994 it was approved for use by the Food and Drug Administration,


The Summer Institute in Statistical Genetics.
The Elizabeth W. Jones Award for Excellence in Education recognizes an individual or group that has had significant, sustained impact on genetics education at any level, from K-12 through graduate school and beyond. Bruce Weir (University of Washington) is the 2019 recipient in recognition of his work training thousands of researchers in the rigorous use of statistical analysis methods for genetic and genomic data. His contributions fall into three categories: the acclaimed Summer Institute in Statistical Genetics, which has been held continuously for 23 years and has trained > 10,000 researchers worldwide; the popular graduate-level textbook Genetic Data Analysis; and the training of a growing number of forensic geneticists during the rise of DNA evidence in courts around the world.


Social media as a tool for detecting underdiagnosed parasitic infections: the case of spirocercosis.
Spirocercosis is a parasitic disease caused by the nematode Spirocerca lupi that affects several canids around the world. In Costa Rica, few spirocercosis cases have been reported; however, it is suspected that the infection is currently underdiagnosed. As seen during the COVID-19 pandemic, social media has sped up the spread of scientific knowledge, raising practioner's awareness of the epidemiology and signs and manifestations of infections. The aim of this work was to use social media as a tool for increasing awareness of spirocercosis among veterinarians and investigate the frequency of cases occurring in Costa Rica. Infographic material about the infection was disseminated by Whatsapp, Facebook, and Instagram from January 2021 to July 2022 using a public account of the Laboratory of Helminthology of the University of Costa Rica. In addition, Google Forms questionnaires were completed by veterinarians to report the number of spirocercosis cases in their clinics, associated manifestations, diagnosis, treatment, and outcome of the disease. Remarkably, since 2020, a six-fold increase in the number of cases per year was noted. Clinical manifestations matched those reported elsewhere, and one of every three dogs infected with S. lupi died spontaneously or required euthanasia because of the infection. This work suggests that S. lupi infection in Costa Rica may have been underdiagnosed in the last decade, and the knowledge and awareness of clinical personnel play a major role in the detection of this and other infectious diseases.


Guided Imagery and Music in the Treatment for Breast and Gynecologic Cancer: An RCT Pilot.
The primary aim of this randomized controlled trial pilot was to investigate the potential effectiveness of the music psychotherapy method, Guided Imagery and Music (GIM), to assist female patients who are undergoing chemotherapy treatment for breast or gynecologic cancer. A secondary aim was to suggest performance improvements regarding the protocol and the resources required to undertake a future larger scale study. The researchers randomized participants (n = 20) into two groups: Intervention group (n = 10) and Control group (n = 10). The Intervention group received a series of six individual, short GIM sessions whereas the Control group received two verbal counseling sessions that took place at Week 1 and at Week 6 of treatment. All participants in the study completed the Profile of Mood States (POMS-Brief); Cancer Fatigue Scale (CFS); and Herth Hope Index (HHI) before the beginning of Week 1 and again after Week 6 or, in the case of the Intervention group, after the last GIM session. All participants also completed two Visual Analogue Scales (VAS-Hope; VAS-Fatigue) weekly or after each session throughout the duration of the trial. The results of the Intervention group receiving GIM showed medium pre-post effect sizes for the CFS, HHI, and POMS questionnaires, and significant positive changes for the VAS-H and VAS-F questionnaires. As seen from this initial data outcome, a brief series of GIM sessions shows promise to be beneficial for increasing hope, decreasing fatigue, and mitigating distressed mood for female patients undergoing treatment for breast or gynecologic cancer.


Impact of increasing prevalence of minimally invasive prostatectomy on open prostatectomy observed in the national inpatient sample and national surgical quality improvement program.
Laparoscopic and especially robot-assisted minimally invasive prostatectomy (MIP) has increased in popularity over the past decade. We analyzed how the increasing prevalence of MIP has affected the outcomes of MIP and open radical prostatectomy (RRP). In the Nationwide Inpatient Sample, 23,473 patients undergoing MIP and 118,266 undergoing RRP between 2002 and 2008 are reported. We analyzed length of stay (LOS), hospital charges (THC), complication rates (CR), and socioeconomic characteristics. We used the American College of Surgeons National Surgical Quality Improvement Program (NSQIP) to identify complication rates (RRP n=666, and MIP n=2205). The proportion of MIP increased from 1.4% in 2002 to 29.5% in 2008. Mean LOS decreased for MIP (2.4 days in 2002, 1.6 days in 2008) and RRP (3.1 days in 2002, 2.1 days in 2008). Mean THC for MIP decreased ($46k in 2002, $34k in 2008) and increased for RRP ($18k in 2002, $32k in 2008). After 2005, overall CRs of MIP were lower than for RRP. High-volume centers reported lower CRs for both procedures. MIP was associated with fewer transfusions and wound complications. Men living in ZIP codes with the top quartile of yearly mean household income were more likely to undergo MIP than RRP (P<0.001). Although there were more white patients receiving MIP and black or Hispanic patients more frequently underwent RRP, there was no statistically significant difference. Increasing use of MIP led to decreased hospital stay for all patients, increase charges for RRP, and decreased CRs for both MIP and RRP. In recent years, MIP was associated with fewer complications. Charges for RRP have increased over time to approach those for MIP, and patients with increased socio-economic status were more likely to undergo MIP.


Finite Element Study on the Preservation of Normal Knee Kinematics with Respect to the Prosthetic Design in Patient-Specific Medial Unicompartmental Knee Arthroplasty.
Alterations in native knee kinematics in medial unicompartmental knee arthroplasty (UKA) are caused by the nonanatomic articular surface of conventional implants. Technology for an anatomy mimetic patient-specific (PS) UKA has been introduced. However, there have been no studies on evaluating the preservation of native knee kinematics with respect to different prosthetic designs in PS UKA. The purpose of this study was to evaluate the preservation of native knee kinematics with respect to different UKA designs using a computational simulation. We evaluated three different UKA designs: a nonconforming design, an anatomy mimetic design, and a conforming design for use under gait and squat loading conditions. The results show that the anatomy mimetic UKA design achieves closer kinematics to those of a native knee compared to the other two UKA designs under such conditions. The anatomy memetic UKA design exhibited a 0.39 mm and 0.36° decrease in the translation and rotation, respectively, in the swing phase compared with those of the natural knee. In addition, under the gait and squat loading conditions, the conforming UKA design shows limited kinematics compared to the nonconforming UKA design. Our results show that the conformity of each component in PS UKA is an important factor in knee joint kinematics; however, the anatomy mimetic UKA design cannot restore perfect native kinematics.


The validity and inter-device variability of the Apple Watch™ for measuring maximal heart rate.
Maximal heart rate (HRmax) is a fundamental measure used in exercise prescription. The Apple Watch™ measures heart rate yet the validity and inter-device variability of the device for measuring HRmax are unknown. Fifteen participants completed a maximal oxygen uptake test while wearing an Apple Watch™ on each wrist. Criterion HRmax was measured using a Polar T31™ chest strap. There were good to very good correlations between the watches and criterion (left: r = 0.87 [90%CI: 0.67 to 0.95]; right: r = 0.98 [90%CI: 0.94 to 0.99]). Standardised mean bias for the left and right watches compared to the criterion were 0.14 (90%CI: -0.12 to 0.39; trivial) and 0.04 (90%CI: -0.07 to 0.15; trivial). Standardised typical error of the estimate for the left and right watches compared to the criterion were 0.51 (90%CI: 0.38 to 0.80; moderate) and 0.22 (90%CI: 0.16 to 0.34; small). Inter-device standardised typical error was 0.46 (90%CI: 0.36 to 0.68; moderate), ICC = 0.84 (90%CI: 0.65 to 0.93). The Apple Watch™ has good to very good criterion validity for measuring HRmax, with no substantial under- or over-estimation. There were moderate and small prediction errors for the left and right watches. Inter-device variability in HRmax is moderate.


Comparison of chemotherapy and hematopoietic stem cell transplantation pre and postterm DMFT scores: a preliminary study.
Chemotherapy is frequently used as a conditioning regimen to destroy malignant marrow cells before transplantation. Xerostomia, dysphagia, altered taste perception, mucositis, soft-tissue ulceration, and infection are common adverse oral effects of chemotherapy. The study was aimed to compare decayed, missing, filled teeth (DMFT) scores before and after hematopoietic stem cell transplantation (HSCT) and chemotherapy. Thirty-six patients undergoing HSCT were included in the study. A pre-HSCT dental treatment protocol was implemented that consisted of restoration of all active carious lesions, treatment of periodontal infections, and extraction of all teeth with advanced periodontal disease. Upon completion of dental treatment, the importance of rigorous and effective oral hygiene was reemphasized, and patients were recalled 6 months later. DMFT scores were calculated prior to the initiation of HSCT treatment and 6 months after transplantation. Regression analysis was used to evaluate the effects of HSCT and chemotherapy on DMFT scores. Wilcoxon T test showed a statistically significant difference in DMFT scores before and after HSCT ( P < 0.001). DMFT scores were found to increase after chemotherapy and HSCT, suggesting that the risk of infection is higher among HSCT patients when compared to other individuals. The results emphasize the need for dental examinations as an integral part of examination and treatment planning for patients undergoing HSCT and chemotherapy.


A novel GmFAD3-2a mutant allele developed through TILLING reduces α-linolenic acid content in soybean seed oil.
Soybean (Glycine max (L.) Merr.) oil typically contains 8% α-linolenic acid that is highly unstable and easily oxidized. This property is undesirable in many food and industrial applications. Genetic strategies for reducing α-linolenic acid content would enhance the commercial value. However, genetic resources for low α-linolenic acid content are limited among natural soybean variations. Microsomal omega-3-fatty acid desaturase (FAD3) is responsible for the synthesis of α-linolenic acid in the polyunsaturated fatty acid pathway. There are four FAD3 homologs (Glyma02g39230, Glyma11g27190, Glyma14g37350 and Glyma18g06950) in the soybean genome. While non-functional alleles have been reported for Glyma02g39230 (GmFAD3-1a) and Glyma14g37350 (GmFAD3-1b), little variation is seen in Glyma18g06950 (GmFAD3-2a). We isolated seven mutant GmFAD3-2a alleles, each containing a single-nucleotide substitution, from 39,100 independent mutant lines by using targeting induced local lesions in genomes (TILLING). Analysis of GmFAD3-2a transcripts and enzyme activities revealed that one missense mutant, 'P1-A9', contains a non-functional allele of GmFAD3-2a. By combining three non-functional alleles (GmFAD3-1a, GmFAD3-1b, and GmFAD3-2a), we generated soybean lines containing <2% α-linolenic acid in their seeds. The reverse-genetics-based development of novel mutant alleles in the fatty acid metabolic pathway will allow the improvement of soybean with better oil quality through conventional breeding.


Design of dendritic macromolecules containing folate or methotrexate residues.
Polyether dendritic compounds bearing folate residues on their surface were prepared as model drug carriers with potential tumor cell specificity. Starting from ester-terminated polyether dendrimers, hydrazide groups were easily introduced to the surface of the dendrimers by reaction with hydrazine. Folate residues were then conjugated to the hydrazide chain ends of the dendrimers by direct condensation with folic acid in the presence of a condensing agent or by reaction with an active ester derivative of folic acid. Essentially complete functionalization of the terminal hydrazide groups was achieved for both the first and the second generation dendrimers with four and eight hydrazide groups. For the G-2 dendrimer with 16 hydrazide groups, an average number of only 12.6 folate residues were attached to each dendrimer. The conjugates are soluble in aqueous medium above pH 7.4. In addition, a similar conjugation of the antitumor drug methotrexate to the dendrimer was also investigated. Once optimized, these molecules may form the basis for a novel family of multivalent drug carriers.


Prevention of rhinovirus and poliovirus uncoating by WIN 51711, a new antiviral drug.
WIN 51711, a potent new antipicornavirus drug, has been shown to inhibit an early event in the replication cycle of human poliovirus type 2 and human rhinovirus type 2. WIN 51711 was not virucidal and had no measurable effect on the adsorption of [3H]uridine-labeled virions to cells. When virion penetration of the plasma membrane was determined through loss of sensitivity to neutralizing antisera, WIN 51711 had no effect on poliovirus penetration, but inhibited rhinovirus penetration by 40%. In the presence of WIN 51711, exposure of neutral red-encapsidated virus-infected cells to light at 3 h postinfection resulted in a 3-log reduction in the number of infectious centers, indicating that WIN 51711 maintained the viral RNA in the encapsidated state after penetrating the cell membrane. The inhibition of uncoating by WIN 51711 in the neutral red assay was found to be concentration dependent, with a concentration of 0.03 micrograms/ml resulting in a 90% inhibition of uncoating. Sucrose gradient sedimentation of lysates from whole cells infected with [3H]uridine-labeled poliovirus showed that poliovirions remained intact in the presence of WIN 51711, but were uncoated in the absence of drug. WIN 51711 also prevented thermal inactivation of poliovirus infectivity, indicating a direct stabilizing effect of this compound on virion capsid conformation.


Muscle carnitine palmityltransferase deficiency and myoglobinuria.
Muscle carnitine palmityltransferase activity, measured by three different methods, was very low (0 to 20 percent of controls) in a patient with a familial syndrome of recurrent myoglobinuria. Long-chain fatty acyl CoA synthetase activity was normal; acetylcarnitine transferase activity was decreased by 40 percent, and carnitine content was 1.7 times higher than the mean control value. Utilization of palmitate by isolated mitochondria was more impaired than utilization of palmitylcarnitine, suggesting a more severe defect of carnitine palmityltransferase I than transferase II. Thus, myoglobinuria may be due to a genetic defect of lipid metabolism in skeletal muscle.


30 Years of Experience with Non-Hodgkin Lymphoma in Children and Adolescents: a retrospective cohort study.
Describe the clinical and demographic characteristics of pediatric patients with non-Hodgkin's lymphoma (NHL) enrolled in a tertiary unit of Pediatric Hematology between 1982-2015. A retrospective cohort study of 140 patients aged 16 years or less with NHL. Demographic characteristics, data on diagnosis, and outcomes were analyzed. The overall survival (OS) analysis and stratification by the most frequent histological subtypes were performed using the Kaplan-Meier method. One hundred and thirty-six patients with de novo NHL and four with NHL as a second malignancy were analyzed. The median age at diagnosis was 6.4 years (interquartile range, 4.2 to 11.1 years); 101 patients were males. Four patients had primary immunodeficiency, four had human immunodeficiency virus, two post-liver transplantation, and one had autoimmune lymphoproliferative syndrome. The most frequent histological type was NHL of mature B- cell (B-NHL-B; 67.1%), with Burkitt's lymphoma being the most frequent subtype, and lymphoblastic lymphoma (LBL, 21.4%). The main clinical manifestation at the diagnosis was abdominal tumors (41.4%). During the follow-up time, 13 patients relapsed, but five of them reached a second remission. Thirty-five patients died, and 103 remained alive in clinical remission. No contact was possible for two patients. The OS at 5 years was 74.5% (± 3.8%). The OS estimated for patients with LBL, NHL-B, and the remaining was 80.4%±7.9%, 72.8%±4.7%, and 74.5%±11%, respectively (P = 0.58). Our results are comparable with cohorts from other middle-income countries.


Method optimization for proteomic analysis of soybean leaf: Improvements in identification of new and low-abundance proteins.
The most critical step in any proteomic study is protein extraction and sample preparation. Better solubilization increases the separation and resolution of gels, allowing identification of a higher number of proteins and more accurate quantitation of differences in gene expression. Despite the existence of published results for the optimization of proteomic analyses of soybean seeds, no comparable data are available for proteomic studies of soybean leaf tissue. In this work we have tested the effects of modification of a TCA-acetone method on the resolution of 2-DE gels of leaves and roots of soybean. Better focusing was obtained when both mercaptoethanol and dithiothreitol were used in the extraction buffer simultaneously. Increasing the number of washes of TCA precipitated protein with acetone, using a final wash with 80% ethanol and using sonication to ressuspend the pellet increased the number of detected proteins as well the resolution of the 2-DE gels. Using this approach we have constructed a soybean protein map. The major group of identified proteins corresponded to genes of unknown function. The second and third most abundant groups of proteins were composed of photosynthesis and metabolism related genes. The resulting protocol improved protein solubility and gel resolution allowing the identification of 122 soybean leaf proteins, 72 of which were not detected in other published soybean leaf 2-DE gel datasets, including a transcription factor and several signaling proteins.


High-throughput secretomic analysis of single cells to assess functional cellular heterogeneity.
Secreted proteins dictate a range of cellular functions in human health and disease. Because of the high degree of cellular heterogeneity and, more importantly, polyfunctionality of individual cells, there is an unmet need to simultaneously measure an array of proteins from single cells and to rapidly assay a large number of single cells (more than 1000) in parallel. We describe a simple bioanalytical assay platform consisting of a large array of subnanoliter microchambers integrated with high-density antibody barcode microarrays for highly multiplexed protein detection from over a thousand single cells in parallel. This platform has been tested for both cell lines and complex biological samples such as primary cells from patients. We observed distinct heterogeneity among the single cell secretomic signatures that, for the first time, can be directly correlated to the cells' physical behavior such as migration. Compared to the state-of-the-art protein secretion assay such as ELISpot and emerging microtechnology-enabled assays, our approach offers both high throughput and high multiplicity. It also has a number of clinician-friendly features such as ease of operation, low sample consumption, and standardized data analysis, representing a potentially transformative tool for informative monitoring of cellular function and immunity in patients.


Microdialysis studies on 3,4-methylenedioxymethamphetamine-induced dopamine release: effect of dopamine uptake inhibitors.
The effect of dopamine (DA) and serotonin (5-HT) uptake inhibitors on 3,4-methylenedioxymethamphetamine (MDMA)-induced increase in DA efflux was studied using in vivo microdialysis. MDMA was infused directly into the anterolateral striatum via the dialysis probe. The local administration of MDMA produced a dose- and time-dependent increase in the extracellular concentration of DA in the striatum. Peripheral administration of the DA uptake blockers, mazindol (5 mg/kg, i.p.) or GBR 12909 (10 mg/kg, i.p.), produced a slight but significant increase in the extracellular concentration of DA. Moreover, pretreatment with either mazindol or GBR 12909 30 min before the infusion of MDMA (10 microM) significantly attenuated the MDMA-induced increase in the extracellular concentration of DA. Pretreatment with fluoxetine (10 mg/kg, i.p.), a 5-HT uptake blocker, 30 min before the infusion of MDMA produced a slight but significant inhibition of MDMA-induced increase in DA concentration. In contrast, pretreatment with the 5-HT2/1C antagonist, ketanserin (3 mg/kg, i.p.), had no significant effect on the increase in DA concentration produced by the local administration of MDMA. These data are suggestive that MDMA increases the concentration of DA in the striatum, in part, via a carrier-mediated mechanism which is largely independent of its effects on 5-HT release.


Hearing of note: an electrophysiologic and psychoacoustic comparison of pitch discrimination between vocal and instrumental musicians.
Cortical auditory evoked potentials of instrumental musicians suggest that music expertise modifies pitch processing, yet less is known about vocal musicians. Mismatch negativity (MMN) to pitch deviances and difference limen for frequency (DLF) were examined among 61 young adult women, including 20 vocalists, 21 instrumentalists, and 20 nonmusicians. Stimuli were harmonic tone complexes from the mid-female vocal range (C4-G4). MMN was elicited by multideviant paradigm. DLF was obtained by an adaptive psychophysical paradigm. Musicians detected pitch changes earlier and DLFs were 50% smaller than nonmusicians. Both vocal and instrumental musicians possess superior sensory-memory representations for acoustic parameters. Vocal musicians with instrumental training appear to have an auditory neural advantage over instrumental or vocal only musicians. An incidental finding reveals P3a as a sensitive index of music expertise.


Prediction of protein-RNA binding sites by a random forest method with combined features.
Protein-RNA interactions play a key role in a number of biological processes, such as protein synthesis, mRNA processing, mRNA assembly, ribosome function and eukaryotic spliceosomes. As a result, a reliable identification of RNA binding site of a protein is important for functional annotation and site-directed mutagenesis. Accumulated data of experimental protein-RNA interactions reveal that a RNA binding residue with different neighbor amino acids often exhibits different preferences for its RNA partners, which in turn can be assessed by the interacting interdependence of the amino acid fragment and RNA nucleotide. In this work, we propose a novel classification method to identify the RNA binding sites in proteins by combining a new interacting feature (interaction propensity) with other sequence- and structure-based features. Specifically, the interaction propensity represents a binding specificity of a protein residue to the interacting RNA nucleotide by considering its two-side neighborhood in a protein residue triplet. The sequence as well as the structure-based features of the residues are combined together to discriminate the interaction propensity of amino acids with RNA. We predict RNA interacting residues in proteins by implementing a well-built random forest classifier. The experiments show that our method is able to detect the annotated protein-RNA interaction sites in a high accuracy. Our method achieves an accuracy of 84.5%, F-measure of 0.85 and AUC of 0.92 prediction of the RNA binding residues for a dataset containing 205 non-homologous RNA binding proteins, and also outperforms several existing RNA binding residue predictors, such as RNABindR, BindN, RNAProB and PPRint, and some alternative machine learning methods, such as support vector machine, naive Bayes and neural network in the comparison study. Furthermore, we provide some biological insights into the roles of sequences and structures in protein-RNA interactions by both evaluating the importance of features for their contributions in predictive accuracy and analyzing the binding patterns of interacting residues. All the source data and code are available at http://www.aporc.org/doc/wiki/PRNA or http://www.sysbio.ac.cn/datatools.asp email@example.com Supplementary data are available at Bioinformatics online.


Role of protein phosphatase 1 in dephosphorylation of Ebola virus VP30 protein and its targeting for the inhibition of viral transcription.
The filovirus Ebola (EBOV) causes the most severe hemorrhagic fever known. The EBOV RNA-dependent polymerase complex includes a filovirus-specific VP30, which is critical for the transcriptional but not replication activity of EBOV polymerase; to support transcription, VP30 must be in a dephosphorylated form. Here we show that EBOV VP30 is phosphorylated not only at the N-terminal serine clusters identified previously but also at the threonine residues at positions 143 and 146. We also show that host cell protein phosphatase 1 (PP1) controls VP30 dephosphorylation because expression of a PP1-binding peptide cdNIPP1 increased VP30 phosphorylation. Moreover, targeting PP1 mRNA by shRNA resulted in the overexpression of SIPP1, a cytoplasm-shuttling regulatory subunit of PP1, and increased EBOV transcription, suggesting that cytoplasmic accumulation of PP1 induces EBOV transcription. Furthermore, we developed a small molecule compound, 1E7-03, that targeted a non-catalytic site of PP1 and increased VP30 dephosphorylation. The compound inhibited the transcription but increased replication of the viral genome and completely suppressed replication of EBOV in cultured cells. Finally, mutations of Thr(143) and Thr(146) of VP30 significantly inhibited EBOV transcription and strongly induced VP30 phosphorylation in the N-terminal Ser residues 29-46, suggesting a novel mechanism of regulation of VP30 phosphorylation. Our findings suggest that targeting PP1 with small molecules is a feasible approach to achieve dysregulation of the EBOV polymerase activity. This novel approach may be used for the development of antivirals against EBOV and other filovirus species.


The International Society for Sexual Medicine's Process of Care for the Assessment and Management of Testosterone Deficiency in Adult Men.
In 2014, the International Society for Sexual Medicine (ISSM) convened a panel of experts to develop an evidence-based process of care for the diagnosis and management of testosterone deficiency (TD) in adult men. The panel considered the definition, epidemiology, etiology, physiologic effects, diagnosis, assessment and treatment of TD. It also considered the treatment of TD in special populations and commented on contemporary controversies about testosterone replacement therapy, cardiovascular risk and prostate cancer. The aim was to develop clearly worded, practical, evidenced-based recommendations for the diagnosis and treatment of diagnosis and management of TD for clinicians without expertise in endocrinology, such as physicians in family medicine and general urology practice. A comprehensive literature review was performed, followed by a structured, 3-day panel meeting and 6-month panel consultation process using electronic communication. The final guideline was compiled from reports by individual panel members on areas reflecting their special expertise, and then agreed by all through an iterative process. This article contains the report of the ISSM TD Process of Care Committee. It offers a definition of TD and recommendations for assessment and treatment in different populations. Finally, best practice treatment recommendations are presented to guide clinicians, both familiar and unfamiliar with TD. Development of a process of care is an evolutionary process that continually reviews data and incorporates the best new research. We expect that ongoing research will lead to new insights into the pathophysiology of TD, as well as new, efficacious and safe treatments. We recommend that this process of care be reevaluated and updated by the ISSM in 4 years.


Nf2/Merlin controls progenitor homeostasis and tumorigenesis in the liver.
The molecular signals that control the maintenance and activation of liver stem/progenitor cells are poorly understood, and the role of liver progenitor cells in hepatic tumorigenesis is unclear. We report here that liver-specific deletion of the neurofibromatosis type 2 (Nf2) tumor suppressor gene in the developing or adult mouse specifically yields a dramatic, progressive expansion of progenitor cells throughout the liver without affecting differentiated hepatocytes. All surviving mice eventually developed both cholangiocellular and hepatocellular carcinoma, suggesting that Nf2(-/-) progenitors can be a cell of origin for these tumors. Despite the suggested link between Nf2 and the Hpo/Wts/Yki signaling pathway in Drosophila, and recent studies linking the corresponding Mst/Lats/Yap pathway to mammalian liver tumorigenesis, our molecular studies suggest that Merlin is not a major regulator of YAP in liver progenitors, and that the overproliferation of Nf2(-/-) liver progenitors is instead driven by aberrant epidermal growth factor receptor (EGFR) activity. Indeed, pharmacologic inhibition of EGFR blocks the proliferation of Nf2(-/-) liver progenitors in vitro and in vivo, consistent with recent studies indicating that the Nf2-encoded protein Merlin can control the abundance and signaling of membrane receptors such as EGFR. Together, our findings uncover a critical role for Nf2/Merlin in controlling homeostasis of the liver stem cell niche.


Cytotoxicity of two autopolymerized acrylic resins used in orthodontics.
To test in vitro the null hypothesis that the toxic effect of different acrylic resins used in orthodontics cannot be reduced when a prior elution is performed. Three established cell lines were used (HeLa, NIH3T3, and Hep2) and cultured under standard conditions. Resin segments were immersed in a culture medium and left to elute for 24 and 48 hours. Cells were exposed to medium containing eluates for 24 or 48 hours. The 3-(4,5 dimethyl-2-thiazolyl)-2.5-diphenyl-2H-tetrazolium bromide (MTT) assay was used as the cytotoxicity test. Control cells contained standard medium with no eluate. Analysis of variance and Tukey test were used for statistical analysis. Fibroblastic viability was not affected when the elution time was 24 hours, but treatments showed higher cell viability than controls when the elution time was 48 hours. When left to elute for 24 hours, both resins had a cytotoxic effect on epithelial cells, but this effect was not observed when the elution time was 48 hours. The hypothesis was rejected as both tested materials showed lower cytotoxic effect when treated with 48-hour elutes compared with 24-hour elutes, which indicates that a longer elution time reduces resin toxicity.


Cranial muscle development in frogs with different developmental modes: direct development versus biphasic development.
Normal development in anurans includes a free swimming larva that goes through metamorphosis to develop into the adult frog. We have investigated cranial muscle development and adult cranial muscle morphology in three different anuran species. Xenopus laevis is obligate aquatic throughout lifetime, Rana(Lithobates) pipiens has an aquatic larvae and a terrestrial adult form, and Eleutherodactylus coqui has direct developing juveniles that hatch from eggs deposited on leaves (terrestrial). The adult morphology shows hardly any differences between the investigated species. Cranial muscle development of E. coqui shows many similarities and only few differences to the development of Rana (Lithobates) and Xenopus. The differences are missing muscles of the branchial arches (which disappear during metamorphosis of biphasic anurans) and a few heterochronic changes. The development of the mandibular arch (adductor mandibulae) and hyoid arch (depressor mandibulae) muscles is similar to that observed in Xenopus and Rana (Lithobates), although the first appearance of these muscles displays a midmetamorphic pattern in E. coqui. We show that the mix of characters observed in E. coqui indicates that the larval stage is not completely lost even without a free swimming larval stage. Cryptic metamorphosis is the process in which morphological changes in the larva/embryo take place that are not as obvious as in normal metamorphosing anurans with a clear biphasic lifestyle. During cryptic metamorphosis, a normal adult frog develops, indicating that the majority of developmental mechanisms towards the functional adult cranial muscles are preserved.


Statistical shape modeling-based algorithm for replacing missing beats in blood pressure signals.
In clinical environments, such as the Intensive Care Unit (ICU), continuous and uninterrupted monitoring of vital signs is critical for the early detection of patient deterioration and prompt intervention. Since data collected in these settings are often corrupted by noise, artifacts, or recording gaps, it is important to estimate missing data for a more accurate signal assessment.In this study, we propose an automatic algorithm for reconstructing of arterial blood pressure signal waveforms. The methodological core of the algorithm is based on the idea of statistical shape modeling, which basically estimates the shape variation of beat waveforms in order to reconstruct them in noisy segments. The waveform reconstruction is achieved by combining the average beat template from a 90-second segment of clean signal preceding the gap with the main shape variations of the estimated waveform.The algorithm was validated using arterial blood pressure recordings from 9 subjects admitted in the ICU and collected in the MIMIC-III Waveform Database, each lasting 1 hour and sampled at 125 Hz. For each record, ten fictitious gaps were created, and the reconstructed segments were compared to the original signals with the metrics proposed within the PhysioNet / Computing in Cardiology Challenge 2010. Results demonstrate the excellent performance of the proposed algorithm, with overall averages of both Q1 and Q2 metrics greater than 0.85.


Curing the Megaplasmid pTT27 from Thermus thermophilus HB27 and Maintaining Exogenous Plasmids in the Plasmid-Free Strain.
Stepwise deletions in the only plasmid in Thermus thermophilus HB27, megaplasmid pTT27, showed that two distantly located loci were important for maintenance of the plasmid. One is a minimum replicon including one gene, repT, coding a replication initiator, and the other encodes subunits of class I ribonucleotide reductase (RNR) for deoxynucleoside triphosphate (dNTP) synthesis. Since the initiator protein, RepT, bound to direct repeats downstream from its own gene, it was speculated that a more-downstream A+T-rich region, which was critical for replication ability, could be unwound for replication initiation. On the other hand, the class I RNR is not necessarily essential for cell growth, as evidenced by the generation of the plasmid-free strain by the loss of pTT27. However, the plasmid-free strain culture has fewer viable cells than the wild-type culture, probably due to a dNTP pool imbalance in the cell. This is because of the introduction of the class I RNR genes or the supplementation of 5'-deoxyadenosylcobalamin, which stimulated class II RNR encoded in the chromosome, resolved the decrease in the number of viable cells in the plasmid-free strain. Likewise, these treatments dramatically enhanced the efficiency of transformation by exogenous plasmids and the stability of the plasmids in the strain. Therefore, the class I RNR would enable the stable maintenance of plasmids, including pTT27, as a result of genome replication normalized by reversing the dNTP pool imbalance. The generation of this plasmid-free strain with great natural competence and its analysis in regard to exogenous plasmid maintenance will expand the availability of HB27 for thermophilic cell factories.


Melanotic macules and melanoacanthomas of the lip. A comparative study with census of the basal melanocyte population.
Fifteen labial melanotic macules and three melanoacanthomas of the lip were studied with particular regard to their distinguishing histopathological features including a quantitative study of the basal melanocyte population. Both of these entities show an increased population of melanocytes arranged as single units along the junctional zone. These conditions differ from each other primarily by the presence of intraepithelial melanocytes in melanoacanthoma. In our opinion, the melanotic macule of the lip has numerous histological counterparts that occur on other mucosal and cutaneous surfaces. Oral mucosal analogies to the labial melanoacanthoma also exist.


Recent update of gastrointestinal endoscope reprocessing.
As infection-related issues have become one of the most important concerns in endoscopy centers, proper reprocessing of endoscopes has attracted great interest. Compliance with established guidelines for reprocessing is critical to prevent pathogen transmission. However, hospital compliance with guidelines has not been satisfactory. To increase compliance, efforts have focused on developing new and more innovative disinfectants and an automated endoscope reprocessor. Reprocessing must be performed by appropriately trained personnel and regular monitoring of reprocessing is essential for quality assurance to improve compliance.


Probing the Dynamics of Streptococcus pyogenes Cas9 Endonuclease Bound to the sgRNA Complex Using Hydrogen-Deuterium Exchange Mass Spectrometry.
The Cas9 endonuclease is an essential component of the CRISPR-Cas-based genome editing tools. The attainment of high specificity and efficiency of Cas9 during targetted DNA cleavage is the main problem that limits the clinical application of the CRISPR-Cas9 system. A deep understanding of the Cas9 mechanism and its structural-functional relationships is required to develop strategies for precise gene editing. Here, we present the first attempt to describe the solution structure of Cas9 from S. pyogenes using hydrogen-deuterium exchange mass spectrometry (HDX-MS) coupled to molecular dynamics simulations. HDX data revealed multiple protein regions with deuterium uptake levels varying from low to high. By analysing the difference in relative deuterium uptake by apoCas9 and its complex with sgRNA, we identified peptides involved in the complex formation and possible changes in the protein conformation. The REC3 domain was shown to undergo the most prominent conformational change upon enzyme-RNA interactions. Detection of the HDX in two forms of the enzyme provided detailed information about changes in the Cas9 structure induced by sgRNA binding and quantified the extent of the changes. The study demonstrates the practical utility of HDX-MS for the elucidation of mechanistic aspects of Cas9 functioning.


Evolutionary trends and ecological differentiation in early Cenozoic Fagaceae of western North America.
• The early Cenozoic was a key period of evolutionary radiation in Fagaceae. The common notion is that species thriving in the modern summer-dry climate of California originated in climates with ample summer rain during the Paleogene.• We investigated in situ and dispersed pollen of Fagaceae from the uppermost Eocene Florissant fossil beds, Colorado, United States, using a combined light and scanning electron microscopy approach.• Pollen types of Castaneoideae with affinities to modern Castanea, Lithocarpus, and Castanopsis were recognized. Pollen of the extinct genus Fagopsis represents a derived type of Castaneoideae pollen. Infrageneric groups of Quercus were well represented, including pollen of Group Protobalanus. The taxonomic diversity of Fagaceae and of the total plant assemblage indicates a mosaic of microclimates, that range from pronounced to weakly seasonal climates and depend on slope aspect and elevation. Continental climatic conditions may have triggered the evolution of sclerophyllous leaves and adaptive radiation in Quercus and other taxa thriving today under distinctly summer-dry and winter-dry climates.• Vegetation types similar to modern vegetation belts of the Coastal Ranges (chaparral, nemoral conifer forest) were established in the Front Range in the late Eocene. Coeval plant assemblages from the Coastal Ranges of California indicate distinctly subtropical, moist climates. Hence, characteristic elements found today in the summer-dry and winter-dry climates of Pacific North America (Quercus Group Protobalanus, Notholithocarpus) may opportunistically have dispersed into their modern ranges later in the Cenozoic. This scenario is in contrast to the evolution and migration patterns of their western Eurasian Mediterranean counterparts (Quercus Group Ilex).


Relationship between individual payload weight and spondylolysis incidence in Turkish land forces.
OBJECTIVEThe aim of this study was to investigate the relationship between lumbar spondylolysis and payload weight between different combat units of Turkish land forces (TLF).METHODThe authors reviewed clinical and radiological data of the military personnel with low-back pain (LBP) admitted to their clinic between July 2017 and July 2018. Age, BMI, average payload weight, and military service unit were recorded. CT scans were evaluated for pars interarticularis fractures and spondylolisthesis, whereas MRI studies were evaluated for spondylolisthesis, Modic-type endplate changes, or signal loss on T2-weighted images compatible with disc degeneration.RESULTFollowing exclusion, a total of 642 all-male military personnel were included. Of these personnel, 122 were commandos, 435 were infantry, and 85 were serving in the artillery units. Bilateral pars interarticularis fracture was noted in 42 commandos (34.42%) and 2 infantrymen (0.45%). There was no spondylolysis in the artillery units. There was no multiple-level spondylolysis and the most common level of spondylolysis was L5. Commandos had a significantly higher incidence of spondylolysis and more average payload weight (p < 0.001). Twelve patients (27.2%) with spondylolysis had accompanying MRI pathologies at the same level, whereas 32 patients (72.7%) had no accompanying MRI pathologies.CONCLUSIONSIncreased payload weight in military personnel is associated with spondylolysis, and commandos in the TLF have significantly heavier payloads, which causes an increased rate of spondylolysis compared to other units. Additionally, spondylolysis without adjacent-level changes on MRI could be undiagnosed. LBP in active military personnel who have a history of carrying heavy payloads should be evaluated extensively with both MRI and CT scans.


Factors underlying prefrontal and insula structural alterations in smokers.
Based upon previous reports of alterations in white matter integrity and gray matter density in smokers, we examined these markers in a large, well-matched sample of smokers and non-smokers. We further investigated the effect of heavy cigarette exposure by using pack-years and the effects of two relatively stable, highly heritable traits in smokers (Fagerström Test of Nicotine Dependence (FTND), a measure of severity of nicotine dependence and Toronto Alexithymia Scale (TAS-20), a stable personality trait related to smoking). Forty-eight nicotine-dependent subjects and 48 matched controls were included in the analyses, with smokers also subdivided into high/low dependence and high/low pack-years smokers. White matter integrity (fractional anisotropy (FA)) and gray matter density (voxel-based morphometry (VBM)) were measured and compared across groups. Gray matter density was lower in left prefrontal cortex (PFC) in high pack-years smokers and was inversely related to pack-years. In contrast, left insular cortex gray matter density was higher in smokers and associated with TAS-20 total score and with difficulty-identifying-feelings factor. Further, the most highly dependent smokers showed lower prefrontal FA, which was negatively correlated with FTND. There was no correlation between pack-years and FTND in our smoker population. These data suggest chronic tobacco use is correlated with prefrontal gray matter damage , while differences in insula gray matter and PFC white matter appear to reflect stable and heritable differences between smokers and non-smokers.


[Effect of vitamin D3 and 20-hydroxyecdysone on the content of ATP, creatine phosphate, carnosine and Ca2+ in skeletal muscles].
The effect of vitamin D3, 20-hydroxyecdysone and extract from Serratula coronata containing 20-hydroxyecdysone on the level of basic metabolites in the skeletal muscles of rats has been studied. It was shown that development of D-hypovitaminosis is accompanied by the decrease in content of ATP, creatine phosphate, carnosine, and by the increase of Ca2+ content. Against the background of D-hypovitaminosis the 20-hydroxyecdysone and the extract from Serratula coronata which contains 20-hydroxyecdysone promote the increase of the amount of these metabolites up to the control of one and normalize Ca2+ content in them.


A water soluble light activated hydrogen sulfide donor induced by an excited state meta effect.
We have utilized an m-amino benzyl based photoremovable protecting group (PRPG) to develop a new water soluble H2S donor. It efficiently releases H2S on demand in a spatio-temporally controlled fashion by an excited state "meta effect" with good chemical and photochemical quantum yield in an aqueous environment. The efficient photorelease of H2S under physiological conditions was also demonstrated by in vitro studies.